Diehl Otávio J, Assano Patrícia K, da Costa Thiago Roncini G, Oliveira Rhaul, Marques-Souza Henrique, Umbuzeiro Gisela de A
School of Technology, University of Campinas, Limeira, 13484-332, Brazil.
Institute of Biology, University of Campinas, Campinas, 13083-862, Brazil.
Ecotoxicology. 2021 May;30(4):751-755. doi: 10.1007/s10646-021-02395-5. Epub 2021 Mar 26.
Regeneration is a widely spread process across the animal kingdom, including many species of marine crustaceans. It is strongly linked to hormonal cycles and, therefore, a great endpoint candidate for toxicology studies. We selected the amphipod Parhyale hawaiensis as test organism, already used in ecotoxicological studies and able to regenerate its body appendages. We are proposing a protocol to use the antenna regeneration as a toxicity endpoint. First, we evaluated differences in time of completion of regeneration in males and females after the amputation of one antenna of 6 months old animals. Then we compared the influence of different testing volumes in the regeneration process (100 and 5 mL). We used as testing substances, dimethyl sulfoxide (DMSO) and diflubenzuron, a chitin synthesis inhibitor. The most suitable protocol consisted of volumes of 5 mL in 12-well microplates, with 1 organism per well, 12 organisms per concentration (1:1 females/males) and test time duration of around 5 weeks. DMSO accelerated regeneration time with a NOEC of 0.06%. Diflubenzuron inhibited the time necessary to its completion with a NOEC of 0.32 μg L. We conclude that the Parhyale hawaiensis antenna regeneration protocol proposed here is a potential tool in ecotoxicology, but more studies are required for its validation not only to verify its utility for testing chemicals but also environmental samples.
再生是动物界广泛存在的过程，包括许多海洋甲壳类物种。它与激素周期密切相关，因此是毒理学研究的一个很好的终点指标。我们选择了夏威夷寻常钩虾作为受试生物，它已被用于生态毒理学研究且能够再生其身体附肢。我们提出了一种将触角再生用作毒性终点指标的方案。首先，我们评估了6月龄动物单侧触角截肢后，雄性和雌性完成再生的时间差异。然后我们比较了不同测试体积（100和5毫升）对再生过程的影响。我们使用二甲基亚砜（DMSO）和几丁质合成抑制剂除虫脲作为测试物质。最合适的方案是在12孔微孔板中使用5毫升的体积，每孔1个生物，每个浓度12个生物（雌雄比例为1:1），测试持续时间约为5周。DMSO加速了再生时间，无可见效应浓度（NOEC）为0.06%。除虫脲抑制了完成再生所需的时间，NOEC为0.32微克/升。我们得出结论，本文提出的夏威夷寻常钩虾触角再生方案是生态毒理学中的一种潜在工具，但需要更多研究来验证它，不仅要验证其用于测试化学品的效用，还要验证其用于环境样品的效用。
